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Mus musculus

ABSTRACT: Next Generation Sequencing Facilitates Quantitative Analysis of Wild Type and Pax9-/- Palate shelves Transcriptomes

PROVIDER: PRJNA352691 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR5000500.fastq.gz	Fastqsanger.gz
	SRR5000501.fastq.gz	Fastqsanger.gz
	SRR5000502.fastq.gz	Fastqsanger.gz
	SRR5000503.fastq.gz	Fastqsanger.gz
	SRR5000504.fastq.gz	Fastqsanger.gz

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Next-Generation Sequencing Facilitates Quantitative Analysis of the Effects of Wnt Agonist Treatments on Palate Formation

Project description:Nonsyndromic clefts of the palate and/or lip are common birth defects arising in about 1/700 live births worldwide. They are caused by multiple genetic and environmental factors, can only be corrected surgically and require complex post-operative care that imposes significant burdens on individuals and society. Our understanding of the molecular networks that control palatogenesis has advanced through studies on mouse genetic models of cleft palate. In particular, the transcription factor Pax9 regulates palatogenesis through the Bmp, Fgf and Shh pathways in mice. But there is still much to learn about Pax9's relationship with other signaling pathways in this process. Here we show alterations of Wnt expression and decreased Wnt activity in Pax9-/- palatal shelves are a likely result of Pax9's ability to directly bind and repress the promoters of Dkk1 and Dkk2, proteins that antagonize Wnt signaling. We exploited this relationship by delivering small-molecule Dkk inhibitors into the tail-veins of pregnant Pax9+/- females from E10.5 to E14.5. Such therapies restored Wnt signaling, promoted cell proliferation, bone formation and fusion of palatal shelves in Pax9-/- embryos. These data uncover a connection between the roles of Pax9 and Wnt genes in palatogenesis and offer a new approach for treating human cleft palates.

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Multimodal transcriptomic profiling of the embryonic palate enables novel in utero cleft therapy [bulk RNA-Seq]

Project description:Cleft palate is a common disorder of development resulting from failure of growth, migration, elevation, and osteogenic fusion of embryonic cranial neural crest-derived palatal shelves. Despite progress in recent decades, the molecular pathways involved in this failure are not well understood. Here, we present a multimodal, spatiotemporal transcriptomic profiling of the developing palate through integrated, unbiased single-cell and bulk RNA-sequencing and multiplexed in situ mRNA mapping of osteogenic cell lineages. We then show that loss of Pax9, a critical transcription factor orchestrator of Wnt signaling in palate development, results in increased expression of sclerostin (Sost), a known antagonist of Wnt signaling. Finally, we reveal that a single dose of sclerostin-neutralizing monoclonal antibody restores Wnt signaling and corrects cleft palate defects in utero in Pax9-/- mouse embryos.

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